Immortalized human brain endothelial cells and flow-based vascular modeling: a marriage of convenience for rational neurovascular studies
Immortalized human brain endothelial cells and flow-based vascular modeling: a marriage of convenience for rational neurovascular studies
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DOI：
10.1038/sj.jcbfm.9600525
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发表时间：
2008-02-01
影响因子：
6.3
通讯作者：
Janigro, Damir
中科院分区：
文献类型：
--
作者：
Cucullo, Luca;Couraud, Pierre-Olivier;Janigro, Damir
在评估进入或排除大脑的药物时，需要新的药物策略。为此，我们开发了一种基于新的人脑血管内皮细胞系（HCMEC/D3）的人源化动态体外血脑屏障模型（hDIV-BBB），其在体内紧密模拟BBB。在该系统中，HCMEC/D3在中空微孔纤维的管腔中生长，并暴露于生理脉动流。与完善的人源化DIV-BBB模型（基于与腔外星形胶质细胞共培养的人脑和非脑血管内皮细胞）的比较证明，即使在不存在腔外星形胶质细胞的情况下，在流动条件下培养的HCMEC/D3细胞也保持了BBB原位的体外生理渗透屏障特性。葡萄糖代谢的测量表明，HCMEC/D3细胞保留有氧代谢途径。对蔗糖和两种相关中枢神经系统药物的渗透性表明，在动态条件下生长的HCMEC/D3细胞密切模仿原位BBB的生理渗透性特性（斜率= 0.93）。还成功地实现了BBB的渗透破坏。DIV-BBB中的峰值BBB开放持续20至30分钟，并且是完全可逆的。此外，在白细胞存在下的血流停止/再灌注序列导致BBB衰竭，如跨内皮电阻的双相降低所示。此外，血脑屏障衰竭是由管腔内释放的促炎因子（白细胞介素-6和白细胞介素-1 β）和基质金属蛋白酶-9（MMP-9）引起的。布洛芬（0.125 mmol/L）预处理可通过降低血流停止/再灌注后的炎症反应来预防BBB衰竭。
In evaluating drugs that enter or are excluded from the brain, novel pharmaceutical strategies are needed. For this reason, we have developed a humanized Dynamic In vitro Blood-Brain Barrier model (hDIV-BBB) based on a novel human brain vascular endothelial cell line (HCMEC/D3), which closely mimics the BBB in vivo. In this system, HCMEC/D3 was grown in the lumen of hollow microporous fibers and exposed to a physiological pulsatile flow. Comparison with well-established humanized DIV-BBB models (based on human brain and non-brain vascular endothelial cells co-cultured with abluminal astrocytes) demonstrated that HCMEC/D3 cells cultured under flow conditions maintain in vitro physiological permeability barrier properties of the BBB in situ even in the absence of abluminal astrocytes. Measurements of glucose metabolism demonstrated that HCMEC/D3 cells retain an aerobic metabolic pathway. Permeability to sucrose and two relevant central nervous system drugs showed that the HCMEC/D3 cells grown under dynamic conditions closely mimic the physiological permeability properties of the BBB in situ (slope = 0.93). Osmotic disruption of the BBB was also successfully achieved. Peak BBB opening in the DIV-BBB lasted from 20 to 30 mins and was completely reversible. Furthermore, the sequence of flow cessation/reperfusion in the presence of leukocytes led to BBB failure as demonstrated by a biphasic decrease in transendothelial electrical resistance. Additionally, BBB failure was paralleled by the intraluminal release of proinflammatory factors (interleukin-6 and interleukin-1 beta) and matrix metalloproteinase-9 (MMP-9). Pretreatment with ibuprofen (0.125 mmol/L) prevented BBB failure by decreasing the inflammatory response after flow cessation/reperfusion.